Multi‐Color, Bleaching‐Resistant Super‐Resolution Optical Fluctuation Imaging with Oligonucleotide‐Based Exchangeable Fluorophores

Super-resolution optical fluctuation imaging (SOFI) is a super-resolution microscopy technique that overcomes the diffraction limit by analyzing intensity fluctuations of statistically independent emitters in time series images. The final images are background-free and show confocality enhanced spatial resolution (super-resolution). Fluorophore photobleaching, however, key limitation for recording long will allow calculation higher order SOFI results with correspondingly increased resolution. Here, we demonstrate photobleaching can be circumvented using fluorophore labels reversibly transiently bind to target, which being replenished from buffer serves as reservoir. Using fluorophore-labeled short DNA oligonucleotides, labeled cellular structures target-specific antibodies contain complementary sequences record events caused transient emitter binding. We this concept bypasses extensive facilitates two-color SOFI.